We attempted to determine the function of genes possibly linked to RNA stability by conducting an experiment with tomato leaf discs and cotyledons. However, we were unable to find any changes in gene expression after cordycepin treatment compared to control (without cordycepin). Also no differences in the gene expression within time points.
Buffer was made up of 1 mm MES, pH 6.25, 1 mm sodium citrate, 1 mm KCl, and 15 mm sucrose) Incubated at 75 rpm for 30 min at room temperature. Cordycepin was added to a final concentration of 0.6 mm. The seedlings or leaf discs were incubated on a shaker at 100 rpm and harvested at 0, 20 minutes, 1h, 3 hrs, 6 hrs, and 9 hrs after treatment with cordycepin.
A new cordycepin was purchased, but the same results were obtained; vacuum infiltration was also tried but did not help.
Appreciate any help,
Thanks
Lola Camalle
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